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United States Patent | 6,258,578 |
Biswas ,   et al. | July 10, 2001 |
The invention provides HIS5 polypeptides and DNA (RNA) encoding HIS5 polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing HIS5 polypeptides to screen for antibacterial compounds.
Inventors: | Biswas; Sanjoy (Paoli, PA); Burnham; Martin Karl Russel (Barto, PA); Lonetto; Michael Arthur (Collegeville, PA); Warren; Patrick Vernon (Philadelphia, PA); Warren; Richard Lloyd (Bluebell, PA) |
Assignee: | SmithKline Beecham Corporation (Philadelphia, PA) |
Appl. No.: | 953139 |
Filed: | October 17, 1997 |
Current U.S. Class: | 435/193; 435/4; 435/6; 435/252.3; 435/320.1; 536/23.2; 536/23.7; 536/24.3; 536/24.32 |
Intern'l Class: | C12N 009/10; C12N 015/00; C12N 001/20; C07H 021/04 |
Field of Search: | 435/193,320.1,252.3,69.1,4,69.7,71.1,91.4,6 536/23.2,23.7,24.3,24.32 |
Foreign Patent Documents | |||
786519 A2 | Jul., 1997 | EP. |
Swissprot Submission, Accession No. Q02132, Direct Submission. (1992). Delorme, C.C. et al. J.Bacteriol. 174:6571-6579 (1992). Berendsen (1998) Science, 282:642-43, 1998.* Database GenBank on STN, Barash et al. (1999) Accession No. V74548.* Ganong (1995) Review of Medical Physiology, Appleton & Lange, Norwalk, Conn.* Galperin, et al., "Sequence Analysis of an Exceptionally Conserved Operon Suggests Enymes for a New Link Between Histidine and Purine Biosynthesis", Molecular Microbiology, vol. 24, No. 2, pp. 443-445, Apr., 1997. Delorme, et al., "Histidine Biosynthesis Genes in Lactococcus lactis subsp. lactis", Journal of Bacteriology, vol. 174, No. 20, pp. 6571-6579, Oct., 1992. Erickson, F. L. and Hannig, E.M., "Characterization of Schizosaccharomyces pombe hisl and his5 cDNAs", Yeast, vol. 11, pp. 157-167, (1995). European Search Report completed Nov. 5, 1999 from corresponding European Application No. 98305979.1, filed Jul. 28, 1998. EP 0 786 519, Esp. pp. 1017-1020, p. 63, pp.1-25, Jul. 30, 1997. EP 0 841 394, Esp. pp. 169-170, pp. 1-17, May 13, 1998. |
TABLE 1 HIS5 Polynucleotide and Polypeptide Sequences (A) Sequences from Staphylococcus aureus HIS5 polynucleotide sequence [SEQ ID NO:1]. 5'-1 ATGATTGTCA TCGTTGATTA TGGATTAGGG AATATTAGTA ATGTAAAACG 51 CGCTATTGAA CATTTAGGGT ATGAGGTGGT TGTCTCAAAT ACCTCAAAAA 101 TAATCGATCA AGCAGAAACA ATCATATTGC CCGGTGTGCGG CCATTTTAAA 151 GATGCGATGT CAGAGATAAA ACGATTAAAT CTCAATGCAA TATTGGCTAA 201 GAATACTGAT AAGAAGATGA TTGGTATTTG TTTAGGCATG CAATTAATGT 251 ATGAGCATAG TGATGAAGGC GATGCATCTG GATTAGGGTT TATCCCAGGA 301 AATATTTCGC GTATCCAAAC AGAATACCCA GTGCCGCACT TAGGTTGGAA 351 TAATTTAGTG AGTAAGCATC CTATGTTAAA TCAAGATGTT TACTTCGTAC 401 ATTCTTACCA AGCGCCGATG TCAGAAAATG TCATTGCATA TGCGCAGTAT 451 GGGGCTGATA TTCCGGCAAT TGTTCAATTT AACAATTATA TTGGTATTCA 501 ATTCCATCCT GAAAAAAGCG GTACATATGG GTTACAAATT TTGCGTCAGG 551 CAATACAAGG GGGATTTATA AATGATTGA -3' (B) HIS5 polypeptide sequence deduced from the polynucleotide sequence in this table [SEQ ID NO:2]. NH.sub.2 -1 MIVIVDYGLG NISNVKRAIE HLGYEVVVSN TSKIIDQAET IILPGVGHFK 51 DAMSEIKRLN LNAILAKNTD KKMIGICLGM QLMYEHSDEG DASGLGFIPG 101 NISRIQTEYP VFHLGWNNLV SKHPMLNQDV YFVHSYQAPM SENVIAYAQY 151 GADIPAIVQF NNYIGIQFHP EKSGTYGLQI LRQAIQGGFI ND-COOH (C) Polynucleotide sequence embodiments [SEQ ID NO:1]. X-(R.sub.1 1).sub.n -1 ATGATTGTCA TCGTTGATTA TGGATTAGGG AATATTAGTA ATGTAAAACG 51 CGCTATTGAA CATTTAGGGT ATGAGGTGGT TGTCTCAAAT ACCTCAAAAA 101 TATTCGATCA AGCAGAAACA ATCATATTGC CCGGTGTCGG CCATTTTAAA 151 GATGCGATGT CAGAGATAAA ACGATTAAAT CTCAATGCAA TATTGGCTAA 201 GAATACTGAT AAGAAGATGA TTGGTATTTG TTTAGGCATG CAATTAATGT 251 ATGAGCATAG TGATGAAGGC GATGCATCTG GATTAGGGTT TATCCCAGGA 301 AATATTTCGC GTATCCAAAC AGAATACCCA GTGCCGCACT TAGGTTGGAA 351 TAATTTAGTG AGTAAGCATC CTATGTTAAA TCAAGATGTT TACTTCGTAC 401 ATTCTTACCA AGCGCCGATG TCAGAAAATG TCATTGCATA TGCGCAGTAT 451 GGGGCTGATA TTCCGGCAAT TGTTCAATTT AACAATTATA TTGGTATTCA 501 ATTCCATCCT GAAAAAAGCG GTACATATGG GTTACAAATT TTGCGTCAGG 551 CAATACAAGG GGGATTTATA AATGATTGA -(R.sub.2).sub.n -Y (D) Polypeptide sequence embodiments [SEQ ID NO:2]. X-(R.sub.1).sub.n -1 MIVIVDYGLG NISNVKRAIE HLGYEVVVSN TSKIIDQAET IILPGVGHFK 51 DAMSEIKRLN LNAILAKNTD KKMIGICLGM QLMYSHSDEG DASGLGFIPG 101 NISRIQTEYP VPHLGWNNLV SKHPMLNQDV YFVHSYQAPM SENVIAYAQY 151 GADIPAIVQF NNYIGIQFHP EKSGTYGLQI LRQAIQGGFI ND-(R.sub.2).sub.n -Y (E) Sequences from Staphylococcus aureus HIS5 polynucleotide ORF sequence [SEQ ID NO:3]. 5'-1 ATGATTGTCA TCGTTGATTA TGGATTAGGG AATATTAGTA ATGTAAAACG 51 CGCTATTGAA CATTTAGGGT ATGAGGTGGT TGTCTCAAAT ACCTCAAAAA 101 TAATCGATCA AGCAGAAACA ATCATATTGC CCGGTGTCGG CCATTTTAAA 151 GATGCGATGT CAGAGATAAA ACGATTAAAT CTCAATGCAA TATTGGCTAA 201 GAATACTGAT AAGAAGATGA TTGGTATTTG TTTAGGCATG CAATTAATGT 251 ATGAGCATAG TGATGAAGGC GATGCATCTG GATTAGGGTT TATCCCAGGA 301 AATATTTCGC TGATCCAAAC AGAATACCCA GTGCCGCACT TAGGTTGGAA 351 TAATTTAGTG AGTAAGCATC CTATGTTAAA TCAAGATGTT TACTTCGTAC 401 ATTCTTACCA AGCGCCGATG TCAGAAAATG TCATTGCATA TGCGCAGTAT 451 GGGGCTGATA TTCCGGCAAT TGTTCAATTT AACAATTATA TTGGTATTCA 501 ATTCCATCCT GAAAAAAGCG GTACATATGG GTTACAAATT TTGCGTCAGG 551 CAATACAAGG GGGATTTATA AATGAT -3' F) HIS5 polypeptide sequence deduced from the polynucleotide ORF sequence in this table [SEQ ID NO:4]. NH.sub.2 -1 MIVIVDYGLG NISNVKRAIE HLGYEVVVSN TSKIIDQAET IILPGVGHFK 51 DAMSEIKRLN LNAILAKNTD KKMIGICLGM QLMYEHSDEG DASGLGFIPG 101 NISRIQTEYP VPHLGWNNLV SKHPMLNQDV YFVHSYQAPM SENVIAYAQY 151 GADIPAIVQF NNYIGIQFHP EKSGTYGLQI LRQAIQGGFI ND-COOH
TABLE 2 Primers for amplification of HIS5 polynucleotides SEQ ID NO PRIMER SEQUENCE 5 5'-ATGTAAAACGCGCTATTGAA-3' 6 5'-CCCCTTGTATTGCCTGACG-3'
SEQUENCE LISTING (1) GENERAL INFORMATION: (iii) NUMBER OF SEQUENCES: 7 (2) INFORMATION FOR SEQ ID NO: 1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 579 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (vi) ORIGINAL SOURCE: (A) ORGANISM: not provided (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1 ATGATTGTCA TCGTTGATTA TGGATTAGGG AATATTAGTA ATGTAAAACG CGCTATTGAA 60 CATTTAGGGT ATGAGGTGGT TGTCTCAAAT ACCTCAAAAA TAATCGATCA AGCAGAAACA 120 ATCATATTGC CCGGTGTCGG CCATTTTAAA GATGCGATGT CAGAGATAAA ACGATTAAAT 180 CTCAATGCAA TATTGGCTAA GAATACTGAT AAGAAGATGA TTGGTATTTG TTTAGGCATG 240 CAATTAATGT ATGAGCATAG TGATGAAGGC GATGCATCTG GATTAGGGTT TATCCCAGGA 300 AATATTTCGC GTATCCAAAC AGAATACCCA GTGCCGCACT TAGGTTGGAA TAATTTAGTG 360 AGTAAGCATC CTATGTTAAA TCAAGATGTT TACTTCGTAC ATTCTTACCA AGCGCCGATG 420 TCAGAAAATG TCATTGCATA TGCGCAGTAT GGGGCTGATA TTCCGGCAAT TGTTCAATTT 480 AACAATTATA TTGGTATTCA ATTCCATCCT GAAAAAAGCG GTACATATGG GTTACAAATT 540 TTGCGTCAGG CAATACAAGG GGGATTTATA AATGATTGA 579 (2) INFORMATION FOR SEQ ID NO: 2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 192 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (vi) ORIGINAL SOURCE: (A) ORGANISM: not provided (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2 Met Ile Val Ile Val Asp Tyr Gly Leu Gly Asn Ile Ser Asn Val Lys 1 5 10 15 Arg Ala Ile Glu His Leu Gly Tyr Glu Val Val Val Ser Asn Thr Ser 20 25 30 Lys Ile Ile Asp Gln Ala Glu Thr Ile Ile Leu Pro Gly Val Gly His 35 40 45 Phe Lys Asp Ala Met Ser Glu Ile Lys Arg Leu Asn Leu Asn Ala Ile 50 55 60 Leu Ala Lys Asn Thr Asp Lys Lys Met Ile Gly Ile Cys Leu Gly Met 65 70 75 80 Gln Leu Met Tyr Glu His Ser Asp Glu Gly Asp Ala Ser Gly Leu Gly 85 90 95 Phe Ile Pro Gly Asn Ile Ser Arg Ile Gln Thr Glu Tyr Pro Val Pro 100 105 110 His Leu Gly Trp Asn Asn Leu Val Ser Lys His Pro Met Leu Asn Gln 115 120 125 Asp Val Tyr Phe Val His Ser Tyr Gln Ala Pro Met Ser Glu Asn Val 130 135 140 Ile Ala Tyr Ala Gln Tyr Gly Ala Asp Ile Pro Ala Ile Val Gln Phe 145 150 155 160 Asn Asn Tyr Ile Gly Ile Gln Phe His Pro Glu Lys Ser Gly Thr Tyr 165 170 175 Gly Leu Gln Ile Leu Arg Gln Ala Ile Gln Gly Gly Phe Ile Asn Asp 180 185 190 (2) INFORMATION FOR SEQ ID NO: 3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 576 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (vi) ORIGINAL SOURCE: (A) ORGANISM: not provided (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3 ATGATTGTCA TCGTTGATTA TGGATTAGGG AATATTAGTA ATGTAAAACG CGCTATTGAA 60 CATTTAGGGT ATGAGGTGGT TGTCTCAAAT ACCTCAAAAA TAATCGATCA AGCAGAAACA 120 ATCATATTGC CCGGTGTCGG CCATTTTAAA GATGCGATGT CAGAGATAAA ACGATTAAAT 180 CTCAATGCAA TATTGGCTAA GAATACTGAT AAGAAGATGA TTGGTATTTG TTTAGGCATG 240 CAATTAATGT ATGAGCATAG TGATGAAGGC GATGCATCTG GATTAGGGTT TATCCCAGGA 300 AATATTTCGC GTATCCAAAC AGAATACCCA GTGCCGCACT TAGGTTGGAA TAATTTAGTG 360 AGTAAGCATC CTATGTTAAA TCAAGATGTT TACTTCGTAC ATTCTTACCA AGCGCCGATG 420 TCAGAAAATG TCATTGCATA TGCGCAGTAT GGGGCTGATA TTCCGGCAAT TGTTCAATTT 480 AACAATTATA TTGGTATTCA ATTCCATCCT GAAAAAAGCG GTACATATGG GTTACAAATT 540 TTGCGTCAGG CAATACAAGG GGGATTTATA AATGAT 576 (2) INFORMATION FOR SEQ ID NO: 4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 192 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (vi) ORIGINAL SOURCE: (A) ORGANISM: not provided (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4 Met Ile Val Ile Val Asp Tyr Gly Leu Gly Asn Ile Ser Asn Val Lys 1 5 10 15 Arg Ala Ile Glu His Leu Gly Tyr Glu Val Val Val Ser Asn Thr Ser 20 25 30 Lys Ile Ile Asp Gln Ala Glu Thr Ile Ile Leu Pro Gly Val Gly His 35 40 45 Phe Lys Asp Ala Met Ser Glu Ile Lys Arg Leu Asn Leu Asn Ala Ile 50 55 60 Leu Ala Lys Asn Thr Asp Lys Lys Met Ile Gly Ile Cys Leu Gly Met 65 70 75 80 Gln Leu Met Tyr Glu His Ser Asp Glu Gly Asp Ala Ser Gly Leu Gly 85 90 95 Phe Ile Pro Gly Asn Ile Ser Arg Ile Gln Thr Glu Tyr Pro Val Pro 100 105 110 His Leu Gly Trp Asn Asn Leu Val Ser Lys His Pro Met Leu Asn Gln 115 120 125 Asp Val Tyr Phe Val His Ser Tyr Gln Ala Pro Met Ser Glu Asn Val 130 135 140 Ile Ala Tyr Ala Gln Tyr Gly Ala Asp Ile Pro Ala Ile Val Gln Phe 145 150 155 160 Asn Asn Tyr Ile Gly Ile Gln Phe His Pro Glu Lys Ser Gly Thr Tyr 165 170 175 Gly Leu Gln Ile Leu Arg Gln Ala Ile Gln Gly Gly Phe Ile Asn Asp 180 185 190 (2) INFORMATION FOR SEQ ID NO: 5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (vi) ORIGINAL SOURCE: (A) ORGANISM: not provided (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5 ATGTAAAACG CGCTATTGAA 20 (2) INFORMATION FOR SEQ ID NO: 6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (vi) ORIGINAL SOURCE: (A) ORGANISM: not provided (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6 CCCCTTGTAT TGCCTGACG 19 (2) INFORMATION FOR SEQ ID NO: 7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 25 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (vi) ORIGINAL SOURCE: (A) ORGANISM: not provided (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7 GCTCCTAAAA GGTTACTCCA CCGGC 25